Characterization and phylogenetic analysis of a Aeromonas veronii clinical strain isolated from blood culture / 中华微生物学和免疫学杂志

Objective To identify an unknown pathogenic strain (X1) isolated from blood culture of an inpatient and to conduct a phylogenetic analysis on it. Methods The X1 strain was isolated from blood culture of an inpatient in the Second Xiangya Hospital of Central South University in 2016. It was identified by morphology biochemical testing and 16S rRNA gene sequencing. Based on genetic distances, a phylogenetic tree was constructed to compare the X1 strain with its homologous species in GenBank in order to reveal its genetic evolution. Results The X1 strain was gram-negative. Biochemical testing for oxidase, glucose and V-P reaction were positive (+) , while the result of citrate was negative (-) . Sequence align-ment analysis of 16S rRNA showed that the X1 strain (1466 bp) was highly homologous with Aeromonas veronii X71120. 1 with a similarity of more than 99％. Genetic distances between the X1 strain and its ho-mologous species, including Aeromonas veronii, Aeromonas hydrophila and Aeromonas sobria, were calculat-ed by MEGA5. 0 and the results revealed that the genetic distance (0. 014) between it and Aeromonas veronii X71120. 1 was the shortest, but the subspecies of it was unable to identify. Cluster analysis also indicated that the X1 strain and Aeromonas veronii were clustered to a small branch. Conclusions The X1 isolate was an Aeromonas veronii strain. Besides, Aeromonas veronii and Aeromonas sobria were similar in genetic char-acteristics.

Effects of expression of growth arrest and DNA damage inducible gene 45α interfered by small interference RNA on the invasion and migration of a cutaneous squamous cell carcinoma cell line A431 / 中华皮肤科杂志

Objective To evaluate effects of targeted silencing of growth arrest and DNA damage inducible gene 45α (Gadd45α) by small interference RNA (siRNA) on the invasion and migration of a cutaneous squamous cell carcinoma cell line A431.Methods Real-time fluorescence-based quantitative PCR and Western blot analysis were performed to detect the mRNA and protein expression of A431 and Colon16 cells respectively,and then A431 cells with highly expressed Gadd45α served as a research object.Cultured A431 cells were divided into 3 groups:experimental group transfected with Gadd45α-siRNA-1,negative control group transfected with negative control siRNA,and blank control group receiving no treatment.After the RNA interference,the mRNA and protein expression of Gadd45α in the above 3 groups were measured by real-time fluorescence-based quantitative PCR and Western blot analysis,respectively,and the effect of the RNA interference on the invasion and migration abilities of A431 cells was evaluated by Transwell and wound scratch assays.Results Gadd45α mRNA and protein were highly expressed in the A431 cells.After the RNA interference,the experimental group showed markedly lower mRNA and protein expressions of Gadd45 in the A431 cells (0.286 ± 0.013,0.33 ± 0.007,respectively) compared with the negative control group (1.028 ± 0.183,0.87 ± 0.002,respectively)and blank control group (1.001 ± 0.057,0.86 ± 0.004,respectively),and there were significant differences in the mRNA and protein expressions of Gadd45 among the 3 groups (F =5 893.857,2 763.000,both P ＜ 0.001).The number of A431 cells crossing the polycarbonate membrane was higher in the experimental group (66.33 ± 3.79) than in the negative control group (26.00 ± 4.36) and the blank control group (28.33 ± 4.16),and there was a significant difference among the 3 groups (F =20.084,P =0.002).Moreover,the wound scratch assay showed that the number of migrating A431 cells per high-power field was significantly higher in the experimental group than in the negative control group and the blank control group (315.33 ± 6.66,154.67 ± 2.08,130.67 ± 3.51 respectively;F =1 676.255,P ＜ 0.001).Conclusion Gadd45α is highly expressed in the cutaneous squamous cell carcinoma cell line A431,and targeted silencing of Gadd45α gene can enhance the in vitro invasion and migration abilities of A431 cells.

In vitro effects of tanshinone ⅡA on autophagy of A375 melanoma cells and related signaling pathway / 中华皮肤科杂志

Objective To investigate in vitro effects of tanshinoneⅡA on the autophagy of A375 melanoma cells and related signaling pathway. Methods Some cultured A375 cells were divided into 5 groups to be treated with tanshinoneⅡA at concentrations of 0.5, 1, 2 and 4 mg/L, and DMEM containing 0.1% dimethyl sulfoxide (DMSO), respectively, for 24, 48, 72 hours. Methyl thiazol tetrazolium (MTT) assay was performed to estimate the proliferative activity of A375 cells. Some cultured A375 cells were divided into 4 groups to be treated with 1, 2 and 4 mg/L tanshinone ⅡA(1?, 2?and 4?mg/L tanshinone group), and DMEM containing 0.1% DMSO (control group), respectively, for 48 hours. Then, flow cytometry was conducted to count autophagosome?positive cells, and Western blot analysis to determine protein expression of autophagy?associated proteins Beclin?1, microtubule?associated protein 1 light chain 3 (LC3)?Ⅱ, phosphatidylinositol 3?kinase(PI3K), protein kinase B(Akt), mammalian target of rapamycin (mTOR)and p70 ribosomal protein S6 kinase 1(p70S6K1). Results MTT assay showed that 24?, 48?, 72?hour treatments with tanshinone ⅡA at concentrations of 0.5, 1, 2 and 4 mg/L all could inhibit the proliferative activity of A375 cells, and the inhibitory effects increased in a dose? and time?dependent manner(F = 2 564.12, 1 235.25, both P < 0.05). The percentage of autophagosome?positive cells and protein expression of Beclin?1 and LC3?Ⅱincreased gradually and significantly in the 1?, 2?and 4?mg/L tanshinone groups(autophagosome?positive cells: 6.91% ± 0.35%, 13.11% ± 0.73%, 25.51% ± 0.83%, respectively; Beclin?1: 0.33 ± 0.01, 0.53 ± 0.04, 0.63 ± 0.02, respectively; LC3?Ⅱ: 0.41 ± 0.01, 0.52 ± 0.02, 0.64 ± 0.02, respectively), after 48?hour treatment, which were significantly different between the tanshinone groups(all P<0.05), and higher in the tanshinone groups than in the control group(0.41%±0.02%;0.09 ± 0.02;0.21 ± 0.01, all P<0.05). However, the protein expression of PI3K, phosphorylated Akt(p?Akt), p?mTOR and p?p70S6K1 in the PI3K?Akt?mTOR?p70S6K1 signaling pathway decreased gradually and significantly with the increase in tanshinone concentrations after 48?hour treatment, and were significantly lower in all the tanshinone groups than in the control group (all P < 0.05). Conclusion Tanshinone ⅡA can promote the auophagy of A375 cells, likely by blocking the PI3K?Akt?mtTOR?p70S6K1 signaling pathway.

Inhibitory effects of tanshinone ⅡA on migration and invasion of melanoma cells and their mechanisms / 中华皮肤科杂志

Objective To evaluate effects of tanshinone ⅡAon the invasion and migration of melanoma A375 cells,as well as on the mRNA and protein expression of CXC chemokine receptor type 7 (CXCR7).Methods In vitro cultured A375 cells were divided into 4 groups to be treated with tanshinone ⅡA at different concentrations of 1,2 and 4 mg/L,and 0.1％ dimethyl sulfoxide (DMSO) (control group) for 48 hours,respectively.Wound scratch assay and Transwell invasion assay were conducted to estimate the migratory and invasive abilities of A375 cells,respectively,and real-time fluorescence-based quantitative PCR (qRT-PCR) and Western blot analysis to determine the mRNA and protein expression of CXCR7 in A375 cells,respectively.Results After 48-hour treatment,the 1-,2-and 4-mg/L tanshinone ⅡA groups showed significantly decreased number of A375 cells crossing the polycarbonate membrane per high-power field (× 200) (71.00 ± 4.00,51.00-± 2.00 and 37.00 ± 3.61,respectively) in Transwell invasion assay,as well as decreased number of A375 cells migrating to the scratch zone (301 ± 3.00,253.00 ± 3.61 and 126.00 ± 7.00,respectively) in wound scratch assay,compared with the control group (105.33 ± 6.51,332.00 ± 6.24,respectively,all P ＜ 0.05).Additionally,qRT-PCR and Western blot analysis revealed that the mRNA and protein expression of CXCR7 was significantly lower in the 1-,2-and 4-mg/L tanshinone ⅡA groups than in the control groups (CXCR7 mRNA:0.63-± 0.04,0.44 ± 0.02 and 0.31 ± 0.01 vs.1.00 ± 0.02;CXCR7 protein:0.573 ± 0.015,0.416 ± 0.011 and 0.260-± 0.055 vs.0.9000 ± 0.010;all P ＜ 0.05).Moreover,inhibitory effects of tanshinone ⅡA on the migration and invasion of A375 cells,as well as on the mRNA and protein expression of CXCR7,were significantly enhanced with the increase of tanshinone ⅡA concentrations (all P ＜ 0.05).Conclusion Tanshinone ⅡA can inhibit the migratory and invasive abilities of melanoma A375 cells by down-regulating CXCR7 expression.

Clinical Observation of Goserelin in the Treatment of Adenomyosis Complicated with Severe Dysmenor-rhea before Placing Levonorgestrel Intrauterine System / 中国药房

OBJECTIVE:To investigate clinical efficacy and safety of goserelin in the treatment of adenomyosis complicated with severe dysmenorrheal before placing levonorgestrel intrauterine system (LIS). METHODS:A total of 140 adenomyosis pa-tients with severe dysmenorrheal were selected from Sanya Family Planning Technology Service Center during Jan. 2014-Jan. 2016, and then divided into control group and observation group according to lottery method,with 70 cases in each group. Both groups re-ceived LIS within first 7 d of menstruation,and observation group was given Goserelin acetate sustained-release implant 10.8 mg subcutaneously via anterior abdominal wall one week before placing. Therapeutic efficacy was evaluated 6 months later. The im-provement of dysmenorrhea was compared between 2 groups. The uterine volume,COX dysmenorrhea score (CMSS),menstrual volume,hemoglobin (Hb) and CA125 content were compared between 2 groups before and after treatment. The occurrence of ADR was also compared. RESULTS:The rate of dysmenorrheal improvement in observation group was 95.71％,which was signifi-cantly higher than 81.43％of control group,with statistical significance(P0.05). Compare to before treatment,CMSS score,menstrual volume and CA125 content of 2 groups were decreased significantly after treatment,while Hb content was increased significantly;the observation group was significantly better than the control group,with statistical significance(P0.05). There was no statistical significance in the incidence of ADR(10.00％ vs. 14.29％)between 2 groups(P>0.05). CONCLUSIONS:Goserelin in the treatment of adenomyosis complicated with severe dysmenorrheal before placing LIS can effectively relieve dysmenorrheal,reduce menstrual volume and improve Hb and CA125 levels,but do not increase the risk of ADR.

Clinical value of serum C-reactive protein, procalcitonin, and lipase in predicting severe acute pancreatitis during pregnancy / 临床肝胆病杂志

ObjectiveTo investigate clinical value of serum C-reactive protein (CRP), procalcitonin (PCT), and lipase in predicting severe acute pancreatitis (SAP) during pregnancy. MethodsA retrospective analysis was performed for the clinical data of 126 pregnant women with acute pancreatitis (AP) who were treated in Nongken Sanya Hospital from January 2011 to January 2016. According to the severity of AP, the patients were divided into control group (pregnancy complicated by mild-to-moderate AP, 71 patients) and observation group (pregnancy complicated by SAP, 55 patients). The clinical features of AP in pregnancy were compared between the two groups. The laboratory markers including venous leukocyte count, hemoglobin, plasma albumin, blood amylase, CRP, PCT, and lipase were observed. The t-test was used for comparision of continuos data between groups, and the chi-square test was used for comparision of categorial data between groups, and the receiver operating characteristic (ROC) curves were plotted for statistically significant markers to assess their sensitivities and specificities. ResultsThe observation group had significantly higher serum levels of lipase, PCT, and CRP than the control group (lipase: 85773±158.61 U/L vs 59019±138.67 U/L, χ2=7.689, P＜0.01; PCT: 10.07±4.55 ng/ml vs 5.89±2.13 ng/ml, χ2=15.492, P＜0.01; CRP: 269.93±63.61 mg/L vs 202.64±39.58 mg/L, χ2=7.500, P＜0.01). According to the ROC curves, serum lipase, PCT, and CRP had areas under the ROC curve of 0.920, 0.841, and 0.832, respectively, with corresponding cut-off values of 712 U/L, 7.6 ng/ml, and 262.0 mg/L, sensitivities of 87.9%, 83.5%, and 81.3%, and specificities of 83.2%, 86.1%, and 78.9%, respectively. ConclusionIncreased serum levels of lipase, PCT, and CRP have a great value in the diagnosis of SAP during pregnancy.

Clinical Observation of Tenofovir Combined with Interferonα1b for Chronic Hepatitis B / 中国药房

OBJECTIVE:To observe clinical efficacy and safety of individual antiviral therapy of tenofovir combined with inter-feron α1b for chronic hepatitis B (CHB). METHODS:96 CHB patients were randomly divided into control group,observation group A and observation group B,with 32 cases in each group. Control group was given entecavir orally,0.5 mg,qd;observation group A was given tenofovir orally,1 piece,qd;observation group B was additionally given interferon α1b,50 μg,3 times a week,on the basis of observation group A. The treatment course lasted for 48 weeks in 3 groups. Clinical efficacy of 3 groups was compared,and the changes of serum liver function indexes,HBV-DNA negative conversion rate and the occurrence of ADR were compared before and after treatment. RESULTS:The total effective rate of observation group B(84.38%)was significantly higher than that of observation group A(62.60%)and control group(37.50%),and that of observation group A was significantly higher than control group,with statistical significance (P0.05). The negative rate of HBV-DNA in observation group B were significantly higher than those in control group and observation group A after 12 and 24 weeks of treatment,with statistical significance(P0.05). No obvious ADR was found in 3 groups. CON-CLUSIONS:Tenofovir combined with interferon α1b shows significant clinical efficacy for CHB,and is significantly better than that of entecavir and tenofovir alone.

Construction and identification of lentiviral vector over-expressing Staphyloco-ccus aureus enterotoxin C3 / 中国免疫学杂志

Objective:To construct the lentiviral vector over-expressing Staphylococcus aureus enterotoxin C3 and detect the expression of target gene in vitro.Methods:SEC3 gene were amplificatied by polymerase chain rcaction( PCR).The GV365 lentiviral vectors were digested by AgeⅠenzyme,which was linked to SEC3 gene and then constructed the GV365-SEC3 lentiviral vetor.Positive clones of vectors were identificd by PCR.Then the positive lentiviral vectors were transfected into 293T cells for lentivirus package.The expression of lentiviral vectors was tested by observating cell fluorescence and Western blot.The virus titer was determined by HIV-1 p24 ELISA.Results: SEC3 gene was amplified and successfully bound to the GV365 lentivirus vectors.The sequences of the recombinant plasmid were confirmed correct by PCR and DNA scqucncing.A large mass of green fluorescent cells were observed after transfecting.And the resulting size of 29 kD protein band of protein electrophoresis, which was consistent with the target gene protein.Viral vector titer was 5×108 TU/ml by ELISA detection.Conclusion: Lentiviral vector over-expressing Staphylococcus aureus enterotoxin C3 was successfully constructed,laid the foundation of observing its effect and mechanism against to tumor in vivo and in vitro for later research.

Effects ofαInterferon on Long-term Survival Rate and Tumor Recurrence After Radical Hepatic Resection / 中国药房

OBJECTIVE:To investigate the effect of α-interferon on long-term survival rate and tumor recurrence after radical hepatic resection. METHODS:104 patients with primary hepatocellular carcinoma underwent radical hepatic resection were divided into observation group(62 cases)and control group(42 cases). Observation group was treated with IFN-α for 18 months 4-6 weeks after surgery;control group received rountine treatment. Total survival rate and disease-free survival rate after surgery were com-pared between 2 groups. RESULTS:In observation group after surgery,1-year survival rate was 82.26%(51/62),3-year survival rate 61.29%(38/62),and 5-year survival rate 24.19%(15/62);in control group after surgery,1-year survival rate was 41.94%(26/62),3-year survival rate 28.57%(12/42)and 5-year survival rate 9.52%(4/42);there was statistically significant difference in postoperative survival rate between 2 groups (P<0.05). In observation group after surgery,1-year disease-free survival rate was 61.29%(38/62),3-year disease-free survival rate 40.32%(25/62),and 5-year disease-free surial rate 17.74%(11/62);in control group after surgery,1 year disease-free survival rate was 43.55%(27/62),3 year disease-free survival rate 22.58%(14/62),and 5-year disease-free surial rate was 0(0/62);there was statistically significant difference in postoperative disease-free survival rate be-tween 2 groups (P<0.05). CONCLUSIONS:The application of α-interferon after radical hepatic resection can significantly inhibit the recurrence of postoperative tumor,improve the long-term survival rate,and is worthy of clinical research.

Imiquimod cream combined with polysaccharide nucleic acid fraction of bacillus Calmette-Guerin for the prevention of recurrent condyloma acuminatum after CO2 laser surgery: an efficacy evaluation / 中华皮肤科杂志

Objective To evaluate the efficacy of imiquimod cream combined with polysaccharide nucleic acid fraction of bacillus Calmette-Guerin (BCG-PSN) for the prevention of recurrent condyloma acuminatum (CA) after CO2 laser surgery.Methods Seventy-four patients with recurrent CA were divided into two groups according to the time of admission and treatment strategies:observation group (n =40) receiving intramuscular BCG-PSN (1.0 mg every other day for 3 months) and applying topical imiquimod cream,control group (n =34) applying topical imiquimod cream only,after CO2 laser surgery.Imiquimod cream was topically used once every other day for 4 weeks in both groups.The percentage of T lymphocyte subsets was determined in these patients before and 3 months after the beginning of treatment.Side effects were recorded during the treatment.All of the patients were followed for 6 months for the evaluation of recurrence.Data were processed with the SPSS 18.0 software,and statistical analysis was carried out by t test and chi-square test.Results After three months of treatment,the patients in the observation group showed a significant increase in the percentage of CD4+ T lymphocytes (42.19％ ± 2.79％ vs.33.56％ ± 3.72％,t =11.74,P＜ 0.05) and CD4/CD8 ratio (1.22 ± 0.21 vs.0.89 ± 0.26,t =6.24,P＜ 0.05),but a significant decrease in the percentage of CD8+T lymphocytes (32.23％ ± 3.32％ vs.39.12％ ± 3.56％,t =8.95,P ＜ 0.05) compared with those before treatment; significant differences were also observed between the observation group and control group in the percentage of CD4+ T lymphocytes (42.19％ ± 2.79％ vs.33.72％ ± 2.94％,t =2.14,P ＜ 0.05) and CD8+ T lymphocytes (32.23％ ± 3.32％ vs.38.47％ ± 3.97％,t =7.26,P ＜ 0.05) as well as CD4/CD8 ratio (1.22 ± 0.21 vs.0.92 ± 0.26,t =5.40,P ＜ 0.05).During the 6 months of follow up,the recurrence rate of CA was 5.00％ (2/40) in the observation group,compared to 47.06％ (16/34) in the control group (x2 =17.66,P ＜ 0.01).The peak of recurrence was observed in the first two months after CO2 laser surgery in both groups,with significant differences between the two groups in the recurrence rate during the first and second months (both P ＜ 0.05).No significant difference was observed in the incidence of adverse reactions between the two groups.Conclusion Imiquimod cream combined with BCG-PSN can effectively enhance immune function and reduce relapse rate in patients with CA after CO2 laser surgery.

Investigation of enterotoxin gene in clinical isolates of Staphylococcus aureus / 南方医科大学学报

<p><b>OBJECTIVE</b>To detect the enterotoxin genes of Staphylococcus aureus (SA) isolated from clinical specimens and analyze the correlation between enterotoxin genes and drug resistance of SA.</p><p><b>METHODS</b>The mecA gene and enterotoxin genes A-F of clinical SA isolates were identified by polymerase chain reaction (PCR), and the genes were sequenced to investigate the correlation of these genes to drug resistance.</p><p><b>RESULTS</b>The detection rate of enterotoxin genes was 100% in 67 methicillin- resistant SA (MRSA), showing no significant difference from the rate in 57 methicillin-sensitive SA (MSSA) (83.5%, χ(2)=0.203, P>0.05). Of the 116 strains carrying enterotoxin genes (93.5%), the detection rates of SEA, SEB, SEC, SED and SEF were 90.5%, 6.9%, 61.3%, 5.2%, 25.9% and 93.5%, respectively, and none of the strains were positive for SEE gene. In these strains, 78 (67.2%) carried 2 or more enterotoxin genes, and the main genotypes were SEA and SEC (33.6%), SEA and SEF (7.8%), and SEA and SEC and SEF (13.8%). Compared with the strains carrying a single enterotoxin gene, those with multiple enterotoxin genes showed a higher drug resistance rate, among which 75% of the SA strains carrying SEA+SEC+SEF were resistant to SXT, significantly higher than the rates of SA carrying SEA (28.6%) and SEA+SEC (38.7%) (P<0.05). The SA strains carrying SEA+SEC+SEF and SEA+SEF showed significantly higher amikacin resistance rates than SA strain carrying SEA (75.0%, 77.0%, 21.5%, respectively, P<0.05).</p><p><b>CONCLUSION</b>Clinical isolates of SA carrying multiple enterotoxin genes have a higher drug resistance rate than those with a single enterotoxin gene, suggesting the the important role of enterotoxin in multidrug resistance.</p>

Polymorphism within the tumor necrosis factor receptor H ( TNFR Ⅱ ) in patients with ankylosing spondylitis in hunan province / 中华微生物学和免疫学杂志

Objective To investigate the association between ankylosing spondylitis (AS) in Han population from Hunan province of China and polymorphism located at the position nt587 in the tumor necrosis factor receptor Ⅱ ( TNFR Ⅱ ) gene.Methods Correlation analysis was performed in a case-control of 100 AS cases and 100 healthy control.The SNPs located in the TNFR Ⅱ gene ( TNFR Ⅱ nt587 ) was examined by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP).With the SPSS software,the relationship between AS and frequencies of genotype and alleles in TNFR Ⅱ nt587 were analyzed.Results There were 43 cases with TNFR Ⅱ nt587 T/T genotype,32 cases with TNFR Ⅱ nt587 T/G genotype and 25 cases with TNFR Ⅱ nt587 G/G genotype in the AS cases.While in the healthy control,there were 56 cases with TNFR Ⅱ nt587 T/T genotype,34 cases with TNFR Ⅱ nt587 T/G genotype and 10 cases with TNFR Ⅱ nt587 G/G genotype.At the same times,the allele frequencies of G in AS group was significantly higher ( x2 =8.734,P=0.003 ) than the control group (41.0％ vs.27.0％ ).The odds ratio in AS cases with TNFR Ⅱ nt587 G/G genotype was 3.256,which higher than those with G/G ( OR =1.226) genotype and T/T genotype compared the healthy control.Conclusion The study demonstrates that polymorphism within the TNFR Ⅱ at the position nt587 is associated with AS and TNFR Ⅱ nt587 G may play an important role in AS susceptibility,and TNFR Ⅱ nt587 G/G genotype may increase the sicken risk of AS in Hunan population.

Study on the gene polymorphism of TNF-α-238 in Ankylosing Spondylitis(AS)patients in Hunan population / 中国免疫学杂志

Objective:To explore the correlations of TNF-α-238 site gene polymorphism and the onset of Ankylosing Spondylitis(AS)in Hunan Han population.Methods:100 AS samples (including serum and whole blood) were collected from the Department of Immunology and Rheumatology of the Second Xiangya hospital from May 2008 to Jan 2009 and 90 samples of normal people were collected as control group.We detected the TNF-α-238 gene polymorphism of these subjects by using PCR-RFLP technique.The TNF-α level in the serum samples were measured by ELISA and HLA-B27 antigen was detected by flow cytometry (FCM).Then all of the data was analyzed by SPSS13.0 software.Results:There were 95 cases with the TNF-α-238 G/G genotype in 100 AS patients,5 cases with the G/A genotype.While in the control group,TNF-α-238 G/G genotype and G/A genotype were 88 cases and 2 cases respectively.There was no TNF-α-238 A/A genotype in both groups.The allele frequencies of G in AS group was higher than in the control group(98.9% vs.97.5%),while the allele frequencies of A in AS group was lower than in the control group(1.1% vs.2.5%).However,there were no significant difference both A allele frequencies and G allele frequencies (P＞0.05).In addition,the average TNF-α level in AS group was higher than in the control group significantly (10.16±1.19 pg/ml vs.5.64±1.18 pg/ml).And the average TNF-α level in AS patients with the genotype of G/A was higher than that of with the G/G genotype (13.49±1.27 pg/ml vs.9.44±1.29 pg/ml).There was a very large difference of the positive ratio of HLA-B27 between two groups (χ~2=114.975,P=0.000).After gene analysis of HLA-B27and TNF-α-238,the odds ratio(OR)was higher in both G/G genotype and HLA-B27 positive than HLA-B27 lonely.Conclusion:There is probably no relationship between the gene polymorphism of TNF-α-238 site and the onset of AS but the G/G genotype of TNF-α-238 may increase the sicken risk of AS in Hunan population.

Biopanning of HCV antigen epitopes from a random 12-peptide library by anti-hepatitis C virus polyclonal antibody / 中南大学学报(医学版)

Objective To analyze the epitopes of anti-hepatitis C virus(HCV)antibodies by peptide library biopanning. Methods Phage random peptide library of 12 amino acids was immunoscreened with purified IgG from the sera of hepatitis C patients. Positive clones which were obtained after 3 rounds of biopanning were detected by ELISA and 4 of them were sequenced. Results After 3 rounds of screening, the radio of output to input increased from (4.6×10~(-4))% to (5.3×10~(-2))%, meaning the enrichment was effective. At the third round of screening, all the selected clones proved to specifically react with the sera for immunoscreening. Four positive phage clones were sequenced, which shared a very conservative sequence and was named as C1. Its inserting sequence in the coat protein III was deduced to be GSMSPYVRWYTP, and the positive rate of C1 reacted with 20 cases of HCV patients was 85%.Conclusion The antigen-mimic peptide C1 is successfully screened from 12 random phage peptide library and it has some diagnostic value.

Changes in bone mineral density and microarchitecture with advancing age in the male apolipoprotein E knockout mice / 中华内分泌代谢杂志

Objective To study the changes of microarchitecture, bone mineral density (BMD) , and bone mineral content (BMC) in apolipoprtein E knockout( ApoE-/-) mice. Methods Male ApoE-/- mice at 15, 28, and 40-week of age and sex-age-matched wild-type (WT) mice were involved in the study. The trabecular and cortical bone microarchitecture were assessed by micro-CT( μCT) in the right distal femur. The total body BMD of the left femur was determined by dual-energy X-ray absorptiometry ( DXA). The relationships among BMD, microarchitecture, and BMC were analyzed. Results Compared with WT mice,the advancing age ApoE-/- mice showed an increased volumetric BMD (vBMD), tissue BMD (tBMD) , BMC, bone volume fraction (BV/TV), trabecular number (Tb. N ) , trabecular thickness (Tb. Th) with an decreased bone surface fraction ( BS/BV), trabecular separation (Tb. SP) , and the structure mode index (P <0. 05 ) in the cancellous bone of femur. The cortical bone microarchitecture parameters as inner perimeter, outer perimeter, cortical area, marrow area, total area and moment of inertia were also increased, but cortical BMD, cortical bone mineral content (C. BMC) and cortical thickness retained constant. At the age of 28 weeks,the total body BMD in ApoEE-/- mice revealed higher than WT mice (P<0. 05) and there was no changes in 15 and 40-week-old mice compared with the sex-age-matched controls. vBMD was positively correlated with BMC, BV/TV,Tb. Th, BS/BV, and C. BMC, with the correlation coefficients 0.955,0.944,0. 834,0.923, and 0.903 .respectively, and there was no correlation between vBMD and the other parameters. Conclusions ApoEE-/- mice display an increased bone mass, suggesting that ApoE has an important role in bone remodeling.

Comparison of phenotype,genotype and drug resistance genes of Acinetobacter baumannii isolated from clinic / 中华微生物学和免疫学杂志

Objective To investigate the genotypes and encoding resistance genes differences of Acinetobacter baumannii and analyze their interrelations with multi-drug resistance.Methods A total of 77strains Acinetobacter baumannii were collected random from the second Xiangya Hospital during September 2008 to September 2009.The K-B method which was WHO recommended was adopted to Acinetobacter baumannii drug sensitivity test to 15 kinds of antibiotics to establish susceptibility spectrum.At the same time,random amplified polymorphic DNA(RAPD)technique was used to establish DNA fingerprinting.The genes of β-lactamase(TEM-1,IMP,OXA-23,OXA-24,AmpC),aminoglycoside-modifying enzymes[aac(3')-Ⅰ ,aac(6')-Ⅰ ,ant(3")-Ⅰ]and 16S rRNA methylase(armA,rmtA,rmtB)were detected by PCR and sequenced,and find the relationship between the gene encoding and multi-drug resistance.In addition,we compared the rates of resistance genes of Acinetobacter baumannii and the relations with the genotype and the multi-resistance.Results Thirty-one sensitive strains and 46 multi-drug resistance strains(10 Pan-drug resistances)were isolated.Seventeen types from A to Q were separated using RAPD technique.E genotype widely popular in the ICU was the advantage type in multi-drug resistance strains,and the rate was 47.1%.While the various types scattered in sensitive strains.The positive rates of TEM-1,IMP,OXA-23,OXA-24,AmpC,aac(3')-Ⅰ ,aac(6')-Ⅰ ,ant(3")-Ⅰ ,armA in the multi-drug resistance strains and the sensitive strains were 95.7%,39.1%,84.8% ,54.3%,87.0%,89.1%,84.8%,45.7%,63.0% and 58.1%,9.7%,32.3%,48.4%,48.4%,29.0%,45.2%,12.9%,9.7%,respectively,and there was significant difference except for OXA-24 using the X2 test(P ＜ 0.05).All isolates were negative for rmtA gene and rmtB gene.Drug susceptibility analysis showed that the resistant rate was significantly higher of the strains carrying resistant genes than that of the resistance negative strains.When the strains were resistant to gentamicin and amikacin,the rate of three aminoglycoside genes positive was 34.8%.The trains containing all the measured β-lactamase genes were all resistant strains.Conclusion Compared with the sensitive Acinetobacter baumannii strains,a broad resistance spectrum and a high drug resistance rate were showed in multidrug resistance strains isolated from clinic,which harboring many kinds of β-lactamase genes and aminoglycosides genes with a high separation rate,and the same clone of multiple drug-resistant strains may be transmitted in and among wards.

Identification of active efflux system gene qacA/B in methicillin-resistant Staphylococcus aureus and its significance / 中南大学学报(医学版)

OBJECTIVE@#To detect the active efflux gene qac gene in methicillin-resistant Staphylococcus aureus (MRSA) by hem-nested polymerase chain reaction (PCR) and to learn the carrier condition of qac gene.@*METHODS@#The active efflux gene qacA/B and qacB of 80 strains MRSA isolated from clinical specimens from Aug 2006 to March 2008 were amplified in vitro by hem-nested PCR with the primers designed by computers based on qac information of Genbank, and the PCR fragments were sequenced and analyzed.@*RESULTS@#We detected qacA/B in 19 out of the 80 MRSA strains (23.75%) and qacB in 18 out of the 80 MRSA strains (22.5%). Compared with sequences of qacA (NO.X56628) and qacB(NO.AF535087) in the Genbank, 98% and 97% were identical, respectively.@*CONCLUSION@#The active efflux gene qac gene in MRSA is detected by hem-nested PCR. The proportion of qac gene positive strains is high in clinical practice, which is related to its multi-antibiotic resistance.

Analysis to Characteristic Chinese Medicines for Treating Rheumatoid Arthritis / 国际中医中药杂志

Objective By studying the prescriptions and medicines for treating rheumatoid arthritis (RA) prescribed by modem doctors, we tried to provide new ideas for developing the new medicines for treating rheumatoid arthritis. Methods 340 prescriptions on treating rheumatoid arthritis were collected from core medical periodicals and books published in the recent 20 years. Make a statistical analysis to the medicines in these prescriptions according to TCM etiological and pharmaceutical theories and modern researching foundlings. Results Among all 254 Chinese medicines used for treating RA, 32 medicines belong to worm and vine medicines, being used for 806 times. Conclusion The worm and vine medicines are frequently used in treating RA with good therapeutic effects combined with other kinds of medicines.

Screening of specific serum biomarker of ankylosing spondylitis from a random peptide library / 中华检验医学杂志

short peptide AS1 screened from the phage random peptide library of 12 amino acids has antigenicity and can react with sera of AS patients. These findings indicate that AS1 could be one of candidate molecules of AS-specific serum markers.

The prevalence of 16S rRNA methylase gene armA and drug resistance in Acinetobacter baumannii / 中华微生物学和免疫学杂志

Objective To investigate the prevalence of 16S rRNA methylase gene armA and to analyze their effect on the drug resistance in multi drug-resistant strains of Acinetobacter baumannii . Methods A total of 72 Acinetobacter baumannii isolates were collected from the Second Xiangya Hospital from Jan. 2008 to Dec. 2008. The size of inhibitory zone of these strains to gentamycin, tobramycin and amikacin were determinate using Kirby-Bauer( K-B) method. The 16S rRNA methylase genes armA were detected by PCR. PCR products were purified and sequenced. Then we used randomly amplified polymorphic DNA method (RAPD) genotyping technology for the establishment of DNA fingerprinting. In addition, we compared drug sensitivity test with RAPD technology. Results Twenty isolates of 72 strains were armA positive and the resistance rates of the strains with armA gene to gentamycin, tobramycin, amikacin were 90.0% , 90.0% and 90. 0% , respectivily. armA positive stains were divided into 7 types using RAPD technology. A genotype was the advantage type. Conclusion The study showed that 16S rRNA methylases gene armA was prevalent in Acinetobacter baumannii which could lead to resistant to almost all aminoglycosides at a high level. And the main form of armA gene prevalence in our hospital was the spread of the same clone strain inside and outside of clinic department.